The present invention relates to tetrahydroquinolines, to processes for their preparation and to their use in medicaments.
The publication U.S. Pat. No. 5,169,857-A2 discloses 7-(polysubstituted pyridyl)-6-heptenoates for treating arteriosclerosis, lipoproteinaemia and hyperproteinaemia. Moreover, the preparation of 7-(4-aryl-3-pyridyl)-3,5-dihydroxy-6-heptenoates is described in the publication EP-325 130-A2. Furthermore, the compound 5(6H)-quinolones,3-benzyl-7,8-dihydro-2,7,7-trimethyl-4-phenyl, is known from the publication Khim. Geterotsikl. Soedin. (1967), (6), 1118-1120.
The present invention relates to tetrahydroquinolines of the general formula (I), 
in which
A represents phenyl which is optionally substituted up to 2 times by identical or different substituents from the group consisting of halogen, trifluoromethyl and straight-chain or branched alkyl or alkoxy having in each case up to 3 carbon atoms,
D represents a radical of the formula 
xe2x80x83in which
R5 and R6 together form a carbonyl group (xe2x95x90O), or
R5 represents hydrogen and
R represents halogen or hydroxyl, or
R5 and R6 represent hydrogen,
R7 and R8 are identical or different and represent phenyl, naphthyl, benzothiazolyl, quinolyl, pyrimidyl or pyridyl which are optionally substituted up to 4 times by identical or different substituents from the group consisting of halogen, trifluoromethyl, nitro, cyano, trifluoromethoxy, or by a radical of the formula xe2x80x94SO2xe2x80x94CH3 or xe2x80x94NR9R10,
xe2x80x83in which
R9 and R10 are identical or different and represent hydrogen or straight-chain or branched alkyl having up to 3 carbon atoms,
E represents cycloalkyl having 3 to 6 carbon atoms, or represents straight-chain or branched alkyl having up to 8 carbon atoms,
R1 represents hydroxyl, and
R2 represents hydrogen or represents methyl,
R3 and R4 are identical or different and represent straight-chain or branched alkyl having up to 3 carbon atoms, or
R3 and R4 together form a spiro-linked alkyl chain having 2 to 4 carbon atoms,
and their salts and N-oxides.
The tetrahydroquinolines according to the invention can also be present in the form of their salts. In general, salts with organic or inorganic bases or acids may be mentioned here.
In the context of the present invention, preference is given to physiologically acceptable salts. Physiologically acceptable salts of the compounds according to the invention can be salts of the substances according to the invention with mineral acids, carboxylic acids or sulphonic acids. Particular preference is given, for example, to salts with hydrochloric acid, hydrobromic acid, sulphuric acid, phosphoric acid, methanesulphonic acid, ethanesulphonic acid, toluenesulphonic acid, benzenesulphonic acid, naphthalenedisulphonic acid, acetic acid, propionic acid, lactic acid, tartaric acid, citric acid, fumaric acid, maleic acid or benzoic acid.
Physiologically acceptable salts can also be metal or ammonium salts of the compounds according to the invention which have a free carboxyl group. Particular preference is given, for example, to sodium, potassium, magnesium or calcium salts, and to ammonium salts, which are derived from ammonia, or organic amines, such as, for example, ethylamine, di-or triethylamine, di- or triethanolamine, dicyclohexylamine, dimethylaminoethanol, arginine, lysine, ethylenediamine or 2-phenylethylamine.
The compounds according to the invention can exist in stereoisomeric forms which are either like image and mirror image (enantiomers), or which are not like image and mirror image (diastereomers). The invention relates both to the enantiomers or diastereomers and to their respective mixtures. These mixtures of enantiomers and diastereomers can be separated into the stereoisomerically uniform components in a known manner.
Preference is given to compounds of the general formula (I) according to the invention
in which
A represents phenyl which is optionally substituted up to 2 times by identical or different substituents from the group consisting of fluorine. chlorine, trifluoromethyl, methyl, ethyl, methoxy and ethoxy,
D represents a radical of the formula 
xe2x80x83in which
R5 and R6 together form a carbonyl group (xe2x95x90O), or
R5 represents hydrogen and
R6 represents fluorine or hydroxyl, or
R5 and R6 represent hydrogen,
R7 and R8 are identical or different and represent phenyl, naphthyl, benzothiazolyl, quinolyl, pyrimidyl or pyridyl, which are optionally substituted up to 3 times by identical or different substituents from the group consisting of fluorine, chlorine, bromine, trifluoromethyl, nitro, cyano, trifluoromethoxy, amino or dimethylamino,
E represents cyclopropyl, cyclobutyl, cyclopentyl or represents straight-chain or branched alkyl having up to 6 carbon atoms,
R1 represents hydroxyl, and
R2 represents hydrogen or represents methyl,
R3 and R4 are identical or different and represent methyl, or
R3 and R4 together form a spiro-linked cyclopropyl, cyclobutyl or cyclopentyl ring,
and their salts and N-oxides.
Particular preference is given to compounds of the general formula (I) according to the invention
in which
A represents phenyl which is optionally substituted up to 2 times by identical or different substituents from the group consisting of fluorine, chlorine, trifluoromethyl, methyl, methoxy and ethoxy,
D represents a radical of the formula 
xe2x80x83in which
R5 and R6 together form a carbonyl group (xe2x95x90O), or
R5 represents hydrogen and
R6 represents fluorine or hydroxyl or
R5 and R6 represent hydrogen,
R7 and R8 are identical or different and represent phenyl, naphthyl, benzothiazolyl, quinolyl, pyrimidyl or pyridyl which are optionally substituted up to 3 times by identical or different substituents from the group consisting of fluorine, chlorine, bromine, trifluoromethyl, nitro, cyano, trifluoromethoxy, amino and dimethylamino,
E represents cyclopropyl or cyclopentyl, or represents straight-chain or branched alkyl having up to 3 carbon atoms,
R1 represents hydroxyl and
R2 represents hydrogen or represents methyl,
R3 and R4 are identical or different and represent methyl, or
R3 and R4 together form a spiro-linked cyclopropyl, cyclobutyl or cyclopentyl ring,
and their salts and N-oxides.
Moreover, processes for preparing the compounds of the general formula (I) according to the invention have been found which are characterized in that
[A] in the case that D represents the radical 
in compounds of the general formula (I) 
xe2x80x83in which
A, E, R1, R2, R3 and R4 are as defined above,
the substituent D is synthesized in inert solvents using organometallic reagents in a Grignard, Wittig or organolithium reaction,
or, in the case that D represents the radical of the formula R8xe2x80x94CH2xe2x80x94Oxe2x80x94CH2, in which R8 is as defined above,
[B] either compounds of the general formula (III) 
xe2x80x83in which
A, E, R1, R2, R3 and R4 are as defined above
xe2x80x83are reacted with compounds of the general formula (IV)
R8xe2x80x94CH2xe2x80x94Zxe2x80x83xe2x80x83(IV), 
xe2x80x83in which
R8 is as defined above, and
Z represents halogen, preferably chlorine or bromine,
xe2x80x83in inert solvents, if appropriate in the presence of a base and/or an auxiliary, or
[C] compounds of the general formula (III) are first converted, by reaction with compounds of the general formula (V) 
xe2x80x83in which
R11 represents straight-chain alkyl having up to 4 carbon atoms,
xe2x80x83into the compounds of the general formula (VI) 
xe2x80x83in which
A, E, R1, R2 R3, R4 and R11 are as defined above,
xe2x80x83and subsequently reacted with compounds of the general formula (VII)
Rxe2x80x2xe2x80x94CH2xe2x80x94OHxe2x80x83xe2x80x83(VII), 
xe2x80x83in which
R8 is as defined above,
xe2x80x83and, if appropriate, protective groups are cleaved off, or
[D] in the case of the compounds of the general formula (Ia) 
xe2x80x83in which
A, E, R3, R4, R5 and R6 are as defined above,
xe2x80x83compounds of the general formula (VIII) 
xe2x80x83in which
A, E, R3, R4 and R7 are as defined above,
xe2x80x83are first oxidized to the compounds of the general formula (IX) 
xe2x80x83in which
R3, R4, R7, A and E are as defined above,
xe2x80x83these are, in a next step, converted by asymmetric reduction into the compounds of the general formula (X) 
xe2x80x83in which
R3, R4, R7, A and E are as defined above,
xe2x80x83these are then converted, by introduction of a hydroxyl-protective group, into the compounds of the general formula (XI) 
xe2x80x83in which
R3, R4, R7, A and E are as defined above and
R12 represents a hydroxyl-protective group, preferably a radical of the formula xe2x80x94SiR13R14R15,
xe2x80x83in which
R13, R14 and R15 are identical or different and represent C1-C4-alkyl,
xe2x80x83from which compound, in a subsequent step, the compounds of the general formula (XII) 
xe2x80x83in which
R3, R4, R7, R12, A and E are as defined above,
xe2x80x83are prepared by diastereoselective reduction,
xe2x80x83and subsequently, by introducing the fluorine substituent with fluorinating agents, such as, for example, DAST and SF4 derivatives, the compounds of the general formula (XIII) 
 in which
R3, R4, R7, R12, A and E are as defined above,
xe2x80x83are prepared,
xe2x80x83and the hydroxyl-protective group is subsequently cleaved off by customary methods,
xe2x80x83and, if appropriate, the substituents listed under D, E and/or R1 and R2 are varied or introduced by customary methods.
The processes according to the invention can be illustrated in an exemplary manner by the following schemes: 
Suitable solvents for all processes are ethers such as diethyl ether, dioxane, tetrahydrofuran, glycol dimethyl ether, or hydrocarbons such as benzene, toluene, xylene, hexane, cylcohexane or mineral oil fractions, or halogenated hydrocarbons such as dichloromethane, trichloromethane, carbon tetrachloride, dichloroethylene, trichloroethylene or chlorobenzene, or ethyl acetate, or triethylamine, pyridine, dimethyl sulphoxide, dimethylformamide, hexamethylphosphoric triamide, acetonitrile, acetone or nitromethane. It is also possible to use mixtures of the solvents mentioned. Preference is given to dichloromethane.
Suitable bases for the individual steps are the customary strongly basic compounds. These preferably include organolithium compounds such as, for example, N-butyllithium, sec-butyllithium, tertbutyllithium or phenyllithium, or amides such as, for example, lithium diisopropylamide, sodium amide or potassium amide, or lithium hexamethylsilylamide, or alkali metal hydrides such as sodium hydride or potassium hydride. Particular preference is given to using N-butyllithium, sodium hydride or lithium diisopropylamide.
Suitable for the processes [B] and [C] are furthermore the customary inorganic bases. These preferably include alkali metal hydroxides or alkaline earth metal hydroxides such as, for example, sodium hydroxide, potassium hydroxide or barium hydroxide, or alkali metal carbonates such as sodium carbonate or potassium carbonate or sodium bicarbonate. Particular preference is given to using sodium hydride or potassium hydroxide.
Suitable organometallic reagents are, for example, systems such as the appropriate Wittig reagents, for example Mg/bromobenzotrifluoride, and the appropriate organolithium compounds, for example p-trifluoromethylphenyllithium.
The reductions are generally carried out using reducing agents, preferably those which are suitable for reducing ketones to hydroxyl compounds Particularly suitable here is the reduction with metal hydrides or complex metal hydrides in inert solvents, if appropriate in the presence of a trialkylborane. The reduction is preferably carried out using complex metal hydrides such as, for example, lithium borohydride, sodium borohydride, potassium borohydride, zinc borohydride, lithium trialkylborohydride, diisobutylaluminium hydride or lithium aluminium hydride. The reduction is very particularly preferably carried out using diisobutylaluminium hydride and sodium borohydride.
The reducing agent is generally employed in an amount of from 1 mol to 6 mol, preferably from 1 mol to 4 mol based on 1 mol of the compounds to be reduced.
The reduction generally proceeds in a temperature range of from xe2x88x9278xc2x0 C. to +50xc2x0 C., preferably from xe2x88x9278xc2x0 C. to 0xc2x0 C. in the case of DIBAH, from 0xc2x0 C. to room temperature in the case of NaBH4, particularly preferably at xe2x88x9278xc2x0 C., in each case depending on the choice of the reducing agent and the solvent.
The reduction generally proceeds at atmospheric pressure; however, it is also possible to operate under elevated or reduced pressure.
The hydrogenation is carried out by customary methods using hydrogen in the presence of noble metal catalysts, such as, for example, Pd/C, Pt/C or Raney nickel in one of the abovementioned solvents, preferably in alcohols such as, for example, methanol, ethanol or propanol, in a temperature range of from xe2x88x9220xc2x0 C. to +100xc2x0 C., preferably from 0xc2x0 C. to +50xc2x0 C., under atmospheric pressure or superatmospheric pressure.
The protective group is generally removed in one of the abovementioned alcohols and THF, preferably methanol THF in the presence of hydrochloric acid in a temperature range of from 0xc2x0 C. to 50xc2x0 C., preferably at room temperature, and atmospheric pressure. In particular cases, preference is given to cleaving off the protective group with tetrabutylammonium fluoride (TBAF) in THF.
In the context of the definition given above, hydroxyl-protective group generally represents a protective group from the series: trimethylsilyl, triisopropylsilyl, tert butyl-dimethylsilyl, benzyl, benzyloxycarbonyl, 2-nitrobenzyl, 4nitrobenzyl, tert butyloxycarbonyl, allyloxycarbonyl, 4-methoxybenzyl, 4-methoxybenzyloxycarbonyl, tetrahydropyranyl, formyl, acetyl, trichloroacetyl, 2,2,2-trichloroethoxycarbonyl, methoxyethoxymethyl, [2-(trimethylsilyl)ethoxy]methyl, benzoyl, 4-methylbenzoyl, 4-nitrobenzoyl, 4-fluorobenzoyl, 4-chlorobenzoyl or 4-methoxybenzoyl. Preference is given to tetrahydropyranyl, tert-butyldimethylsilyl and triisopropylsilyl. Particular preference is given to tert-butyldimethylsilyl.
Suitable solvents for the individual steps are ethers such as diethyl ether, dioxane, tetrahydrofuran, glycol dimethyl ether, diisopropyl ether or hydrocarbons such as benzene, toluene, xylene, hexane, cyclohexane or mineral oil fractions, or halogenated hydrocarbons such as dichloromethane, trichloromethane, carbon tetrachloride, dichloroethylene, trichloroethylene or chlorobenzene. It is also possible to use mixtures of the solvents mentioned.
Suitable oxidizing agents for preparing the compounds of the general formula (IX) are, for example, nitric acid, acid, cerium(IV) ammonium nitrate, 2,3-dichloro-5,6-dicyanobenzoquinone, pyridinium chlorochromate (PCC), pyridinium chlorochromate on basic alumina, osmium tetroxide and manganese dioxide. Preference is given to manganese dioxide and nitric acid.
The oxidation is carried out in one of the abovementioned chlorinated hydrocarbons and water. Preference is given to dichloromethane and water.
The oxidizing agent is employed in an amount of from 1 mol to 10 mol, preferably from 2 mol to 5 mol, based on 1 mol of the compounds of the general formula (VIII).
The oxidation generally proceeds at a temperature of from xe2x88x9250xc2x0 C. to +100xc2x0 C., preferably from 0xc2x0 C. to room temperature.
The oxidation generally proceeds at atmospheric pressure. However, it is also possible to carry out the oxidation under elevated or reduced pressure.
The asymmetric reduction to the compounds of the general formula (X) is generally carried out in one of the abovementioned ethers or toluene, preferably tetrahydrofuran and toluene.
The reduction is generally carried out using enantiomerically pure 1R,2S-aminoindanol and borane complexes such as BH3xc3x97THF, BH3xc3x97DMS and BH3xc3x97(C2H5)2NC6H5. Preference is given to the system borane diethylaniline/1R,2S-aminoindanol.
The reducing agent is generally employed in an amount of from 1 mol to 6 mol, preferably from 1 mol to 4 mol based on 1 mol of the compounds to be reduced.
The reduction generally proceeds at a temperature of from xe2x88x9278xc2x0 C. to +50xc2x0 C., preferably from 0xc2x0 C. to 30xc2x0 C.
The reduction generally proceeds at atmospheric pressure; however, it is also possible to operate under elevated or reduced pressure.
The hydroxyl-protective group is introduced in one of the abovementioned hydrocarbons, dimethylformamide or THF, preferably in toluene in the presence of lutidine in a temperature range of from xe2x88x9220xc2x0 C. to +50xc2x0 C., preferably from xe2x88x925xc2x0 C. to room temperature and atmospheric pressure.
Reagents for introducing the silyl protective group are, in general, tert-butyldimethylsilyl chloride or tert-butyldimethylsilyl trifluoromethanesulphonate. Preference is given to tert-butyldimethylsilyl trifluoromethansulphonate.
The reduction to the compounds of the general formula (XII) proceeds in one of the abovementioned hydrocarbons, preferably toluene.
The reduction for preparing the compounds of the general formula (XII) is generally carried out using customary reducing agents, preferably those which are suitable for reducing ketones to hydroxyl compounds are. Particularly suitable for this purpose is the reduction with metal hydrides or complex metal hydroxides in inert solvents, if appropriate in the presence of a trialkylborane. The reduction is preferably carried out using complex metal hydrides such as, for example, lithium borohydride, sodium borohydride, potassium borohydride, zinc borohydride, lithium trialkylborohydride, diisobutylaluminium hydride, sodium bis-(2-methoxyethoxy)-aluminium hydride or lithium aluminium hydride. Very particularly preferably, the reduction is carried out using sodium bis-(2-methoxyethoxy)-aluminium hydride.
The reducing agent is generally employed in an amount of from 1 mol to 6 mol, preferably from 1 mol to 3 mol based on 1 mol of the compounds to be reduced.
The reduction generally proceeds at a temperature of from ⊕20xc2x0 C. to +110xc2x0 C., preferably from 0xc2x0 C. to room temperature.
The reduction generally proceeds at atmospheric pressure; however, it is also possible to operate under elevated or reduced pressure.
In the reduction to the compounds of the general formula (XII), small residues of the wrong diastereomer remain in the mother liquor. These residues can be reoxidized with customary oxidizing agents such as, for example, pyridinium chlorochromate (PCC) or activated manganese dioxide, in particular with activated manganese dioxide to protected (XI) and thus be recycled into the synthesis cycle without loss of yield.
The fluorine substituent is generally introduced in one of the abovementioned hydrocarbons or methylene chloride, preferably in toluene and under an atmosphere of protective gas.
SF4 derivatives are generally understood to be diethylaminosulphur trifluoride (DAST) or 2,2xe2x80x2-bisfluoro-substituted amines such as, for example, diethyl-1,2,3,3,3-hexafluoropropylamine.
The reaction generally proceeds at a temperature of from xe2x88x9278xc2x0 C. to 100xc2x0 C., preferably, in the case of the diethylaminosulphur trifluoride, at from xe2x88x9278xc2x0 C. to RT and, in the case of the diethyl-1,1,2,3,3,3-hexafluoropropylamine, at from room temperature to 80xc2x0 C.
The protective group is generally cleaved off in one of the abovementioned alcohols and THF, preferably methanol/THF in the presence of hydrochloric acid in a temperature range of from 0xc2x0 C. to 50xc2x0 C., preferably at room temperature, and atmospheric pressure. In particular cases, preference is given to cleaving off the protective group with tetrabutylammonium fluoride (TBAF) in TFF at room temperature.
Derivatizations which may be mentioned by way of example are the following reaction types:
oxidations, reductions, hydrogenations, halogenation, Wittig/Grignard reactions and amidations/sulphoamidations.
Suitable bases for the individual steps are the customary strongly basic compounds. These preferably include organolithium compounds such as, for example, n-butyllithium, sec-butyllithium, tertbutyllithium or phenyllithium, or amides such as, for example, lithium diisopropylamide, sodium amide or potassium amide, or lithium hexamethylsilylamide, or alkali metal hydrides such as sodium hydride or potassium hydride. Particular preference is given to using N-butyllithium, sodium hydride or lithium diisopropylamide.
Suitable bases are furthermore the customary inorganic bases. These preferably include alkali metal hydroxides or alkaline earth metal hydroxides such as, for example, sodium hydroxide, potassium hydroxide or barium hydroxide, or alkali metal carbonates such as sodium carbonate or potassium carbonate or sodium bicarbonate. Particular preference is given to using sodium hydroxide or potassium hydroxide.
Suitable solvents for the individual reaction steps are also alcohols such as methanol, ethanol, propanol, butanol or tertbutanol. Preference is given to tertbutanol.
If required, it is necessary to carry out some reaction steps under an atmosphere of protective gas.
The halogenations are generally carried out in one of the abovementioned chlorinated hydrocarbons, and preference is given to methylene chloride.
Suitable halogenating agents are, for example, diethylamino-sulphur trifluoride (DAST), morpholino-sulphur trifluoride or SOCl2.
The halogenation generally proceeds in a temperature range of from xe2x88x9278xc2x0 C. to +50xc2x0 C., preferably from xe2x88x9278xc2x0 C. to 0xc2x0 C., in each case depending on the choice of halogenating agent and solvent.
The halogenation generally proceeds at atmospheric pressure; however, it is also possible to operate under elevated or reduced pressure.
The compounds of the general formulae (II) and (HI) are novel and can be prepared by preparing,
by reacting the compounds of the general formula (XIV) 
xe2x80x83in which
E is as defined above and
R16 represents C1-C4-alkoxycarbonyl or aryl (D=aryl)
with aldehydes of the general formula (XV)
Axe2x80x94CHOxe2x80x83xe2x80x83(XV), 
xe2x80x83in which
A is as defined above,
xe2x80x83and compounds of the general formula (XVI) 
xe2x80x83in which
R17 and R18 together with a carbonyl group have the abovementioned scope of R1 and R2,
xe2x80x83the compounds of the general formula (XVII) 
xe2x80x83in which
A, E, R16, R17and R18 are as defined above
xe2x80x83and, in the case of the compounds of the general formula (E), then carrying out a reduction, as described above, to give the hydroxymethyl function,
xe2x80x83and, in a last step, converting the alkoxycarbonyl group (R16) by a reduction-oxidation sequence into an aldehyde group.
Suitable solvents for the oxidation are ethers such as diethyl ether, dioxane, tetrahydrofuran, glycol dimethyl ether, or hydrocarbons such as benzene, toluene, xylene, hexane, cyclohexane or mineral oil fractions, or halogenated hydrocarbons such as dichloromethane, trichloromethane, carbon tetrachloride, dichloroethylene, trichloroethylene or chlorobenzene, or ethyl acetate, or triethylamine, pyridine, dimethyl sulphoxide, dimethylformamide, hexamethylphosphoric triamide, acetonitrile, acetone or nitromethane. It is also possible to use mixtures of the solvents mentioned. Preference is given to methylene chloride.
Suitable oxidizing agents are, for example, cerium(IV) ammonium nitrate, 2,3-di-chloro-5,6-dicyano-benzoquinone, pyridinium chlorochromate (PCC), pyridinium chlorochromate on basic alumina, osmium tetroxide and manganese dioxide. Preference is given to sulphur trioxide-pyridine complex in DMSO/methylene chloride and pyridinium chlorochromate on basic alumina.
The oxidizing agent is employed in an amount of from 1 mol to 10 mol, preferably from 2 mol to 5 mol, based on 1 mol of the compounds of the general formula (XVII).
The oxidation generally proceeds in a temperature range of from xe2x88x9250xc2x0 C. to +100xc2x0 C., preferably from 0xc2x0 C. to room temperature.
The oxidation generally proceeds at atmospheric pressure. However, it is also possible to carry out the oxidation under elevated or reduced pressure.
The compounds of the general formulae (IV), (V), (VII), (XIV), (XV) and (XVI) are known per se or can be prepared by customary methods.
Some of the compounds of the general formulae (VI) and (XV) are known or novel, in which case they can be prepared as described above.
The compounds of the general formulae (IX) and (X) are novel species and can be prepared as described above.
The compounds of the general formula (VIII) are novel and can be prepared by reacting
compounds of the general formulae (XVa), (XVIII) and (XIX) 
xe2x80x83in which
A, E, R3, R4 and R7 are as defined above,
xe2x80x83with an acid.
Suitable solvents for preparing the compounds of the general formula (VIII) are the abovementioned ethers or alcohols. Preference is given to diisopropyl ether.
Suitable acids for preparing the compounds of the general formula (VI) are, in general, organic carboxylic acids and inorganic acids, such as, for example, oxalic acid, maleic acid, phosphoric acid, fumaric acid and trifluoroacetic acid. Preference is given to trifluoroacetic acid and oxalic acid.
The acid is generally employed in an amount of from 0.1 mol to 5 mol, preferably 1 mol, based on 1 mol of the compounds of the general formula (XIX).
The reaction is generally carried out at atmospheric pressure. However, it is also possible to carry out the reaction under elevated or reduced pressure.
The reaction is generally carried out at the reflux temperature of the solvent in question.
The compounds of the general formulae (XV) and (XIX) are known per se or can be prepared by customary methods.
The compounds of the general formula (XVIII) are novel and can be prepared by first preparing, by reacting the compounds of the general formula (XX)
Exe2x80x94CO2xe2x80x94R19xe2x80x83xe2x80x83(XX) 
in which
E is as defined above and
R19 represents C1-C4-alkyl
with compounds of the general formula (XXI) 
in which
R7 is as defined above
in a solvent in the presence of 18-crown-6 ether, the compounds of the general formula (XXII) 
in which
R7 and E are as defined above,
followed by reaction with ammonium acetate in inert solvents.
Suitable solvents for the first step of the process are the abovementioned ethers and hydrocarbons, and preference is given to tetrahydrofuran.
Suitable solvents for the reaction with the compounds of the general formula (XXII) are alcohols, such as, for example, methanol, ethanol, propanol or isopropanol. Preference is given to ethanol.
All steps of the process are carried out at the respective reflux temperature of the solvent in question and at atmospheric pressure.
Some of the compounds of the general formulae (XX) and (XXI) are known, or they can be prepared by known methods.
Some of the compounds of the general formula (XXII) are novel species, and they can be prepared as described above.
The compounds of the general formulae (I) and (la) according to the invention have a pharmacological activity spectrum which could not have been foreseen.
The compounds of the general formulae (I) and (la) according to the invention have useful pharmacological properties which are superior when compared to the prior art; in particular, they are highly effective inhibitors of cholesterol ester transfer protein (CETP) and they stimulate the reverse cholesterol transport. The active compounds according to the invention effect a reduction of the LDL cholesterol level in the blood and simultaneously increase the HDL cholesterol level. They can therefore be used for the treatment and prevention of hyperlipoproteinaemia, dyslipidaemias, hypertriglyceridaemias, hyperlipidaemias or arteriosclerosis.
The pharmacological activity of the substances according to the invention was assessed using the following test:
CETP Inhibition Test
Preparation of CETP
CETP is obtained in partially purified form from human plasma by differential centrifugation and column chromatography and used for the test. For this purpose, human plasma is adjusted to a density of 1.21 g per ml using NaBr and centrifuged at 50,000 rpm at 4xc2x0 C. for 18 h. The bottom fraction (d greater than 1.21 g/ml) is applied to a Sephadex(copyright)Phenyl-Sepharose 4B (Pharmacia) column, washed with 0.15 m NaCl/0.001 M TrisHCl pH 7.4 and subsequently eluted using dist. water. The CETP-active fractions are pooled, dialysed against 50 mM Na-acetate pH 4.5 and applied to a CM-Sepharose(copyright) (Pharmacia) column. Elution is subsequently carried out using a linear gradient (0-1 M NaCl). The pooled CETP fractions are dialysed against 10 mM TrisHCl pH 7.4 and subsequently purified further by chromatography over a Mono Q(copyright) column (Pharmacia).
Preparation of Radioactively Labelled HDL
50 ml of fresh human EDTA plasma are adjusted to a density of 1.12 using NaBr and centrifuged at 4xc2x0 C. in a Ty 65 rotor at 50,000 rpm for 18 h. The upper phase is used to obtain cold LDL. The lower phase is dialysed against 3*4 l of PDB buffer (10 mM Tris/HCl pH 7.4, 0.15 mM NaCl, 1 mM EDTA, 0.02% NaN3). Per 10 ml volume of retained material, 20 ml of 3H-cholesterol (Dupont NET-725; 1 mC/ml, dissolved in ethanol) are subsequently added, and the mixture is incubated at 37xc2x0 C. under N2 for 72 h.
The mixture is then adjusted to a density of 1.21 using NaBr and centrifuged in a Ty 65 rotor at 20xc2x0 C. and 50,000 rpm for 18 h. The upper phase is collected and the lipoprotein fractions are purified by gradient centrifugation. To this end, the isolated labelled lipoprotein fraction is adjusted to a density of 1.26 using NaBr. In each case 4 ml of this solution are covered in centrifuge tubes (SW 40 rotor) with 4 ml of a solution of a density of 1.21 and 4.5 ml of a solution of 1.063 (density solutions of PDB buffer and NaBr), and the tubes are subsequently centrifuged in an SW 40 rotor at 38,000 rpm and 20xc2x0 C. for 24 h. The intermediate layer which is found between a density of 1.063 and a density of 1.21 and which contains the labelled HDL is dialysed against 3*100 volume of =PDB buffer at 4xc2x0 C.
The retained material contains radioactively labelled 3H-CE-HDL, which is used for the test adjusted to approximately 5xc3x97106 cmp per ml.
CETP Test
To assess the CETP activity, the transfer of 3H-cholesterol ester from human HD-lipoproteins to biotinylated LD-lipoproteins is measured.
The reaction is terminated by addition of Streptavidin-SPA(copyright) beads (Amersham) and the transferred radioactivity is directly measured in a liquid scintillation counter.
In the assay mixture, 10 ml of HDL-3H-cholesterol ester (50,000 cpm) with 10 ml of Biotin-LDL (Amersham) in 50 mM Hepes/0.15 m NaCl/0.1% bovine serum albumin/0.05% NaN3 pH 7.4 are incubated with 10 ml of CETP (1 mg/ml) and 3 ml of a solution of the substance to be tested (dissolved in 10% DMSO/1% BSA) at 37xc2x0 C. for 18 h. 200 ml of the SPA streptavidin bead solution (TRKQ 7005) are subsequently added, the mixture is incubated with shaking for another 1 h and subsequently measured in a scintillation counter.
The controls used are corresponding incubations with 10 ml of buffer, 10 ml of CETP at 4xc2x0 C. and 10 ml of CFRP at 37xc2x0 C.
The activity which is transferred in the control experiments with CETP at 37xc2x0 C. is taken to be 100% transfer. The substance concentration at which this transfer is reduced by half is stated as the IC50 value.
In Table A below, the IC50 values (mol/l) for CETP inhibitors are given:
Ex vivo Activity of the Compounds According to the Invention
Syrian gold hamsters, which have been bred in our own laboratory, are anaesthetized after 24 hours of fasting (0.8 mg/kg of atropine, 0.8 mg/kg of Ketavet(copyright) s.c., 30xe2x80x2 later 50 mg/kg of nembutal i.p.). The jugular vein is subsequently exposed and cannulated. The test substance is dissolved in a suitable solvent (usually adalate placebo solution: 60 g of glycerol, 100 ml of H2O, ad 1000 ml PEG400) and administered to the animals via a PE catheter, which is introduced into the jugular vein. The same volume of solvent without test substance is administered to the control animals. The vein is subsequently tied off and the wound is closed.
The test substances can also be administered p.o. by dissolving the substances in DMSO and suspending them in 0.5% tylose and administering them perorally using a pharyngeal tube. Identical volumes of solvent without test substance are administered to the control animals.
At different intervalsxe2x80x94up to 24 hours after the administrationxe2x80x94blood samples are taken from the animals by puncture of the retro-orbital venous plexus (approximately 250 ml). Coagulation is completed by incubation at 4xc2x0 C. overnight, and the samples are subsequently centrifuged at 6000xc3x97 g for 10 minutes. The CETP activity is determined in the resulting serum using the modified CETP test. The transfer of 3H-cholesterol ester from HD-lipoproteins to biotinylated LD-lipoproteins is measured as described above for the CErP test.
The reaction is terminated by addition of Streptavidin-SPAO beads (Amersham), and the transferred radioactivity is directly determined in a liquid scintlation counter.
The test protocol is carried out as described under xe2x80x9cCETP testxe2x80x9d. However, to test the serum, 10 ml of CETP are replaced by 10 ml of the appropriate serum samples. Corresponding incubations of sera of untreated animals serve as controls.
The activity that is transferred in the control experiments using control sera is classified as 100% transfer. The substance concentration at which this transfer is reduced by half is stated as the ED50 value.
In vivo Activity of the Compounds According to the Invention
In experiments for assessing the oral activity on lipoproteins and triglycerides, test substance, dissolved in DMSO and suspended in 0.5% tylose, is administered perorally using a pharyngeal tube to Syrian gold hamsters which have been bred in our own laboratory. To determine the CETP activity, blood samples (approximately 250 ml) are taken by retro-orbital puncture prior to the start of the experiment. The test substances are subsequently administered perorally using a pharyngeal tube. Identical volumes of solvent without test substance are administered to the control animals. Subsequently, the animals have to fast and at different intervalsxe2x80x94up to 24 hours after the administration of the substancesxe2x80x94blood samples are taken by puncture of the retro-orbital venous plexus.
Coagulation is completed by incubation at 4xc2x0 C. overnight, and the samples are subsequently centrifuged at 6000xc3x97 g for 10 minutes. The content of cholesterol and triglycerides in the resulting serum is assessed using modified commercially available enzyme tests (cholesterol enzymatic 14366 Merck, triglycerides 14364 Merck). Serum is diluted in a suitable manner with physiological saline solution.
100 ml of serum dilution and 100 ml of test substance are transferred into 96-well-plates and incubated at room temperature for 10 minutes. The optical density is subsequently determined at a wavelength of 492 nm using an automatic plate reader. The triglyceride and cholesterol concentrations of the samples are determined with the aid of a standard curve measured in parallel.
The determination of the HDL-cholesterol content is carried out after precipitation of the ApoB-containing lipoproteins using a reagent mixture (Sigma 352-4HDL cholesterol reagent) in accordance with the instructions of the manufacturer.
In vivo Activity in Transgenic hCETP Mice
The substances to be tested were administered to transgenic mice, which were bred in our own laboratory (Dinchuck, Hart, Gonzalez, Karmann, Schmidt, Wirak; BBA (1995), 1295, 301), via the feed. Prior to the beginning of the experiment, blood samples were taken retro-orbitally from the mice to determine cholesterol and triglycerides in the serum. The serum was obtained as described above for hamsters by incubation at 4xc2x0 C. overnight and subsequent centrifugation at 6000xc3x97 g. After one week, blood samples were again taken from the mice to determine lipoproteins and triglycerides. The change of the measured parameters are expressed as a change in percent based on the initial value.
The invention furthermore relates to the combination of tetrahydroquinolenes of the general formula (I) with a glucosidase and/or amylase inhibitor for the treatment of familial hyperlipidaemias, of obesity (adipositas) and of diabetes mellitus. Glucosidase and/or amylase inhibitors in the context of the present invention are, for example, acarbose, adiposine, voglibose, miglitol, emiglitate, MDL-25637, camiglibose (MDL-73945), tendamistate, AI-3688, trestatin, pradimicin-Q and salbostatin.
Preference is given to the combination of acarbose, miglitol, emiglitate or voglibose with one of the abovementioned compounds of the general formula (I) according to the invention.
Furthermore, the compounds according to the invention can be combined with cholesterol-lowering vastatines or ApoB-lowering principles, in order to treat dyslipidaemi as, combined hyperlipidaemias, hypercholesterolaemias or hypertriglyceridaemias.
The abovementioned combinations can also be used for primary or secondary prevention of coronary heart diseases (for example myocardial infarction).
Vastatins in the context of the present invention are, for example, lovastatin, simvastatin, pravastatin, fluvastatin, atorvastatin and cerivastatin. ApoB-lowering agents are, for example, MTP inhibitors.
Preference is given to the combination of cerivastatin or ApoB inhibitors with one of the abovementioned compounds of the general formula (I) according to the invention.
The novel active compounds can be converted in a known manner into the customary formulations, such as tablets, coated tablets, pills, granules, aerosols, syrups, emulsions, suspensions and solutions, using inert, non-toxic, pharmaceutically suitable carriers or solvents. In this case the therapeutically active compound should in each case be present in a concentration from approximately 0.5 to 90% by weight of the total mixture, i.e. in amounts which are sufficient in order to achieve the dosage range indicated.
The formulations are prepared, for example, by extending the active compounds using solvents and/or carriers, if appropriate using emulsifiers and/or dispersants, it optionally being possible, for example, to use organic solvents as auxiliary solvents if the diluent used is water.
Administration is carried out in a customary manner, intravenously, orally, parenterally or perlingually, in particular orally.
In the case of parenteral administration, solutions of the active compound can be used by employing suitable liquid carrier materials.
In general, it has proved advantageous, in the case of intravenous administration, to administer amounts from approximately 0.001 to 1 mg/kg, preferably approximately 0.01 to 0.5 mg/kg, of body weight to achieve effective results, and in the case of oral administration the dosage is approximately 0.01 to 20 mg/kg, preferably 0.1 to 10 mg/kg, of body weight.
In spite of this, if appropriate it may be necessary to depart from the amounts mentioned, namely depending on the body weight or on the type of administration route, on individual reaction towards the medicament, the manner of its formulation and the time at or interval during which administration takes place. Thus, in some cases it may be adequate to manage with less than the abovementioned minimum amounts, while in other cases the upper limit mentioned has to be exceeded. In the case of the administration of relatively large amounts, it may be advisable to divide these into several individual doses over the course of the day.
Abbreviations Used:
Starting Materials